FORMULATION AND EVALUATION OF NAPROXEN PRONIOSOMAL GEL

M. PHARM. DISSERTATION PROTOCOL

Submitted to

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE ,

KARNATAKA

By

Miss. KAVITA B. JEEDI B. Pharm

Under the guidance ofDr. ANITA R. DESAI M.Pharm.,PhD.

Asso. Professor

DEPARTMENT OF PHARMACEUTICSH.S.K. COLLEGE OF PHARMACY

BAGALKOT-587 101

(2012-13)

ANNEXURE IIPROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1 Name of the Candidate and Address KAVITA B. JEEDIH.S.K. COLLEGE OF PHARMACYB.V.V.S CAMPUSBAGALKOT-587101

PERMANENT ADDRESSKAVITA B. JEEDID/O, BASAPPA A. JEEDIMALLIKARJUN SAW MILLAT POST: MUDAGAL-584125TQ: LINGASAGUR, DIST: RAICHURKARNATAKA

2 Name of the Institution H.S.K. COLLEGE OF PHARMACYB.V.V.S CAMPUSBAGALKOT-587101

3 Course of the Study and Subject

M. PHARMACY (PHARMACEUTICS)

4 Date of Admission 02 - JULY - 2012

5 Title of the project:- “FORMULATION AND EVALUATION OF NAPROXEN PRONIOSOMAL GEL”

6.0 BRIEF RESUME OF THE INTENDED WORK:6.1) Need for the study:

Proniosomes are dry formulations which make use of carriers coated with non-

ionic surfactants and can be converted into niosomes immediately before use by hydration.

‘Proniosomes’ minimize problems of niosome physical stability such as aggregation, fusion,

leaking, and provide additional convenience in transportation, distribution, storage and dosing.

Proniosomes-derived niosomes are as good as or even better than conventional niosomes1.

Proniosomes offer a versatile vesicle drug delivery concept with potential for

delivery of drugs via transdermal route. This would be possible if proniosomes form niosomes

upon hydration with water from skin following topical application under occlusive conditions2.

Transdermal therapeutic system have generated an interest as they provide considerable

advantage of a non-invasive parentral route for drug therapy, avoidance of first pass gut and

hepatic metabolism, decreased side effects and relative ease of drug input termination in

problematic cases3.

Colloidal particulate carriers such as liposomes4 or niosomes5 have been

widely employed in drug delivery systems and producing them from proniosomes provides them

a distinctive advantage. These carriers act as drug reservoirs and the rate of drug release can be

controlled by modification of their composition. These lipid vesicles can carry both hydrophilic

drugs (by encapsulation) and hydrophobic drugs (in lipid domain). Due to their capability to carry

a variety of drugs, these lipid vesicles have been extensively used in various drug delivery

systems6 like drug targeting7, controlled release8 and permeation enhancement of drug9.

To get the desired characteristics of a particular proniosome gel formulation, it is

important to select the surfactant of suitable HLB in the formulation of proniosome gel. The

finding of the studies on proniosomes till date opens the door for the future use of different carrier

materials with biocompatibility and suitability for the preparation of proniosomes.

Non-steroidal anti-inflammatory medicines (NSAIM’s) are a class of drugs that

provide analgesic and antipyretic(fever-reducing) effects, and, in higher doses, anti-

inflammatory effects. The most prominent members of this group of drug is naproxen. Most of

NSAID’s act as nonselective inhibitors of the enzyme cyclooxygenase (COX), inhibiting both

the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) isoenzymes COX catalyzes the

formation of prostaglandin and thromboxane from arachidonic acid itself derived from the

cellular phospholipid bilayer by phospholipase A2prostaglandins act (among other things) as

messenger molecules in the process of inflammation. The discovery of COX-2 led to research to

development of selective COX-2 inhibiting drugs that do not cause gastric problems characteristic

of older NSAID’s. Most non-steroidal anti-inflammatory drugs are weak acids, with a pKa of 3-5.

They are absorbed well from the stomach and intestinal mucosa. They are highly protein-bound in

plasma (typically >95%), usually to albumin, so that their volume of distribution typically

approximates to plasma volume. Most NSAID’s are metabolised in the liver by oxidation and

conjugation to inactive metabolites that typically are excreted in the urine, though some drugs are

partially excreted in bile.

Naproxen is propionic acid derivatives of NSAID’s. It is particularly potent in

inhibiting leukocyte migration may be more valuable in acute gout. It is also responsible for

ankylosing spondylitis. Its plasma half is 12- 16 hrs10.

6.2) Review of Literature:

Alsarra IA et al., investigated permeation properties of ketorolac proniosome gel across excised

rabbit skin using Franz diffusion cell. Proniosomes prepared with Span 60 provided a higher

ketorolac flux than Tween 20. A change in cholesterol content and lecithin content did not

significantly decrease the flux. The encapsulation efficiency and size of niosomal vesicles were

characterized by high performance liquid chromatography method and scanning electron

microscopy. The prepared proniosomes achieved about 99% drug encapsulation14.

Chandra A et al., formulated and optimized a piroxicam proniosomes by using surfactant (span

60), cholesterol and lecithin. The results revealed that span 60 based lecithin vesicles showed

significant (p < 0.05) reduction in paw swelling. The percent inhibition was found to be more than

that of piroxicam carbopol gel. It is probable that there is enhanced drug delivery from lipid

vesicles. The short fall seen with maltodextrin and sorbitol based formulations account for the

slow release observed in in-vitro studies15.

Solanki AB et al., studied the preparation, optimization and characterization of ketoprofen

proniosomes by a central, composite Box-Wilson design. Total lipid concentration (X1), surfactant

loading (X2) and amount of drug (X3) were chosen as independent variables. The proniosomes

were characterized for percentage drug entrapment and mean volume diameter. It was observed

that X1, X2 and X3 influenced the release of drug from proniosomal gel. Developed niosomal gel

showed higher permeation compared to plain ketoprofen gel16.

Gupta A et al., developed a proniosomal carrier system for captopril by transdermal drug delivery

system using various ratios of sorbitan fatty acid esters, cholesterol and lecithin prepared by

coacervation-phase separation method. The method of proniosome loading resulted in an

encapsulation yield of 66.7-78.7%. Proniosomal gel possessed high entrapment efficiency and

utilizes alcohol, which itself can act as a penetration enhancer. At refrigerated conditions, higher

drug retention was observed17.

Hengjiu H et al., described procedure for producing a dry product which may be hydrated

immediately before use to yield aqueous niosome dispersions similar to those produced by more

cumbersome conventional methods. This report describes the preparation of dispersions of

proniosome derived niosomes, comparison of these niosomes to conventional niosomes, and

optimization of proniosome formulations. In addition, conventional and proniosome derived

niosomes are compared in terms of their morphology, particle size, particle size distribution, and

drug release performance in synthetic gastric or intestinal fluid. In all comparisons, proniosome

derived niosomes are as good as or better than conventional niosomes. Further they suggest that

these prepared proniosomes minimize problems of niosome physical stability such as aggregation,

fusion and leaking and provide additional convenience in transportation, distribution, storage and

dosing18.

Mahmoud M et al., developed proniosomal gels or solutions of flurbiprofen based on span 20,

span 40, span 60 and span 80 with and without cholesterol. Nonionic surfactant vesicles

(niosomes) formed immediately upon hydrating proniosomal formulae. They studied influence of

different processing and formulation variables such as surfactant chain length, cholesterol content,

drug concentration, total lipid concentration, negatively or positively charging lipids and pH of the

dispersion medium on flurbiprofen percentage encapsulation efficiency and also, they studied

release of the prepared niosomes in phosphate buffer (pH 7.4). Results indicated that the

percentage encapsulation efficiency followed the trend Sp 60 >Sp 40 >Sp 20 >Sp 80.Cholesterol

increased or decreased the percentage encapsulation efficiency depending on either the type of the

surfactant or its concentration within the formulae. The maximum loading efficiency was 94.61%

when the hydrating medium was adjusted to pH 5.5. Increasing total lipid or drug concentration

also increased the percentage encapsulation efficiency of flurbiprofen into niosomes. However,

incorporation of either dicetyl phosphate (DCP) which induces negative charge or stearyl amine

(SA) which induces positive charge decreased the percentage encapsulation efficiency of

flurbiprofen into niosomal vesicles. Finally, they suggest in vitro release data for niosomes of Sp

40 and Sp 60 showed release profiles of flurbiprofen from niosomes of different cholesterol

contents is an apparently biphasic release process. As a result, this study suggested the potential of

proniosomes as stable precursors for the immediate preparation of niosomal carrier systems19.

Ajay S et al., characterize and optimize aceclofenac proniosomes using central composite design

and carry out stability studies. They selected three independent variables molar ratio of drug to

lipid (X1), surfactant loading (X2) and volume of hydration (X3). Based on central composite

design, they prepared 16 batches of proniosomes by slurry method and evaluated for the

percentage drug entrapment and mean volume diameter. The percentage drug entrapment and

mean volume diameter (dependent variables) and the transformed values of independent variables

were subjected to multiple regressions to establish a second order polynomial equation. Contour

7.0

plots were constructed to further elucidate the relationship between the independent and dependent

variables. The conformity of the polynomial equations was checked by preparing three checkpoint

batches. From the computer optimization process and contour plots, predicted levels of

independent variables X1, X2, and X3 (-0.77, -0.8 and 0 respectively), for an optimum response of

percentage drug entrapment with constraints on mean volume diameter were determined. The

optimized batch was subjected to stability studies. The polynomial equations and contour plots

developed using central composite design suggested preparation of proniosomes with optimum

responses20.

6.3 Objectives of the study :The present study is planned with the following objectives:

1.To prepare naproxen proniosome gel using various surfactants.

2.To study drug surfactants interaction by using DSC and FT-IR instruments.

3.To characterize the formulations for various physicochemical parameters.

4.To evaluate the proniosome gel for in vitro diffusion studies using Franz

Diffusion cell.

5.To carry out stability studies for selected formulations as per ICH guidelines.

Materials and Methods :

7.1 Drugs to be used in the formulations:

Drug : Naproxen.

Excipients : Cholesterol, Soya Lecithin, Span 20, 40, 60, 80.

Tween 20, 40, 60, 80.

Method : Development by coacervation phase separation method.

7.2 Source of Data:

Review of Literature from

a) Journals such as

I. Indian Journal of Pharmaceutical Sciences.

II. Indian Drugs.

III. Journal of Controlled Release.

IV. International Journal of Pharmaceutics.

V. Pakistan Journal of Pharmaceutical Sciences.

VI. Drug Development and Industrial Pharmacy.

VII. Acta Pharma.

b) world wide web.

c) J–Gate@helinet.

d) Library : H.S.K College of Pharmacy.

7.3 Method of collection of data:

The data for the study is planned to collect from the laboratory based experiments:

1. Preformulation studies like solubility, melting point and characterization of the drug and

surfactants will be done by employing suitable methods and compatibility of drug with

surfactants will be carried out by using Infra-red spectroscopy and Differential scanning

calorimetry instruments adopting reported methods.

2. Proniosomal gel of naproxen will be prepared by coacervation phase separation method by

using Soya Lecithin, Cholesterol, Span 20, 40, 60, 80 and Tween 20,40,60,80.

3. The prepared gel will be characterized for vesicle size analysis, rate of spontaneity,

encapsulation efficiency, In vitro release studies will be carried out by using Franz

diffusion cell and the drug release data will be subjected to various kinetic models and

stability studies of selected formulations will be carried out using stability chamber as per

ICH guidelines.

VII.4 Does the study require any investigations or interventions to be conducted on patients or

other humans or animals? If so, Please describe briefly.

-----Not applicable-----

7.5 Has ethical clearance been obtained from your Institution in case of 7.3?

-----Not applicable-----

List of References :

8.01. Walve JR, Rane BR, Gujrathi NA, Bakaliwal SR, Pawar SP. Proniosomes: A surrogated

carrier for improved transdermal drug delivery system. Int J Res Ayu Pharm 2011; 2(3):

743-750.

2. Fang JY, Yu SY, Wu PC, Huang YB, Tsai YH. In-vitro skin permeation of estradiol from

various proniosomes formulation. Int J Pharm 2001; 215:91-99.

3. Wu PC, Huang YB, Chang JJF, Chang JS, Tsai YH. Evaluation of pharmacokinetics and

pharmacodynamics of captopril from transdermal hydrophilic gel in normotensive rabbit

and spontaneously hypertensive rats. Int J Pharm 2000; 209: 87-94.

4. Biju SS, Talegaonkar S, Misra PR, Khar RK. Vesicular systems: An overview. Ind J

Pharm Sci 2006; 68:141-153.

5. Shahiwala A, Misra A. Studies in topical application of niosomally entrapped nimesulide. J

Pharm Sci 2002; 5(3):220-225.

6. Puglia C, Trombetta D, Venuti V, Saija A, Bonina F. Evaluation of in vivo topical anti-

inflammatory activity of indometacin from liposomal vesicles. J Pharm Pharmacol 2004;

56: 1225-1232.

7. Gupta PN, Mishra V, Singh P, Rawat A, Dubey P, Mahor S, Vyas SP. Tetanus toxoid

loaded transfersomes for topical immunization. J Pharm Pharmacol 2005; 57: 295-301.

8. Barber R, Shek P. Liposomes as a topical ocular drug delivery system in Pharmaceutical

particulate carriers. A. Rolland (Ed.), Marcel Dekker, NY 1993; 1–20.

9. Verma DD, Verma S, Blume G, Fahr A. Liposomes increase skin penetration of entrapped

and non-entrapped hydrophilic substances into human skin: a skin penetration and confocal

laser scanning microscopy study. Eur J Pharm Biopharm 2003; 55: 271-277.

10. Tripathi KD, Essentials of medical pharmacology 5th ed, jaypee brothers medical publisher,

2004

11. Muller RH, Radtke M, Wissing SA. Solid lipid nanoparticles (SLN) and nano-structured lipid

carriers (NLC) in cosmetic and dermatological preparations. Adv Drug deliv Rev

2002;54:131-155.

12. Baillie A, Florence A, Hume L, Muirhead G,Rogerson A. Preparation and properties of

niosomes-nonionic surfactant vesicles. J Pharm Pharmacol 1985;37:863-868.

13. Yoscioka T, Sternberg B, Florence T. Preparation and properties of vesicles (niosomes)

of sorbitan monoesters (Span 20,40,60 & 80) and a sorbitan triester (Span 85). Int J

Pharm 1994;105:1-6.

14. Alsarra IA, Bosela AA, Ahmed SM, Mahrous GM. Proniosomes as a drug carrier for

transdermal delivery of ketorolac. Eur J Pharm and Bio 2005; 59: 485–490.

15. Chandra A, Sharma PK. Proniosome based drug delivery system of piroxicam. Afr J

Pharm Pharmacol 2008; 2(9):184-190.

16. Solanki AB, Parikh JR, Parikh RH. Preparation, optimization and characterization of

ketoprofen proniosomes for transdermal delivery. Int J PharmSci Nanotech 2009;

2(1):413-420.

17. Gupta A, Prajapati SK, Balamurugan M, Singh M, Bhatia D. Design and development of a

proniosomal transdermal drug delivery system for captopril tro J Pharm Res 2007;6(2):

687-693.

18. Hengjiu H, David GR. Proniosomes: A novel drug carrier preparation. Int J Pharm

2000;206:110-122.

19. Mahmoud M, Omaima AS, Mohammed AH, Nagia AM. Effect of some formulation

parameters on flurbiprofen encapsulation and release rates of niosomes prepared from

proniosomes. Int J Pharm 2008 ;361:104-111.

20. Ajay S, Jolly P, Rajesh P. Preparation, characterization, optimization, and stability studies

of aceclofenac proniosomes. Ira J Pharm Res 2008;7(4):237-246

21. Cappello B, Maio CD, Iervolino M, Miro A, Improvement of solubility and stability of

valsartan by hydroxyl propyl-betacyclodextrin J Inclusion Phenom Macrocyclic chem,

2006;54:289-294.

22. Schreier H, Bouwstra J, Liposomes and niosomes as topical drug carriers, dermal and

transdermal drug delivery. J Control Rel 1994;30:1-15.

9 Signature of the Candidate (KAVITA B. JEEDI)

10 Remarks of the Guide: The topic selected for dissertation is satisfactory

and feasible

11 Name and Designation :

11.1. Guide: Dr. ANITA R. DESAI M.Pharm.,Ph.D. ASSO. PROFFESSORHEAD OF THE DEPARTMENT OFPHARMACEUTICSH.S.K COLLEGE OF PHARMACY,BAGALKOT-587101

11.2. Signature of Guide (Dr. ANITA R. DESAI)

11.3. Co-Guide NOT APPLICABLE

11.4. Signature of Co- Guide NOT APPLICABLE

11.5. Head of the Department: Dr. ANITA R. DESAI M.Pharm.,Ph.D.

ASSO. PROFFESSORHEAD OF THE DEPARTMENT OFPHARMACEUTICSH.S.K. COLLEGE OF PHARMACY,BAGALKOT-587101

11.6. Signature of HOD(Dr. ANITA R. DESAI)

12 12.1. Remarks of the principal

All the required facilities will be provided to carry out dissertation work under the supervision of the guide.

12.2. PrincipalDr. I. S. MUCHCHANDI M.Pharm., Ph.D.

PRINCIPALH.S.K. COLLEGE OF PHARMACYBAGALKOT-587101

12.3. Signature of the Principal(Dr. I. S. MUCHCHANDI)